Plate Heights below 50 nm for Protein Electrochromatography Using Silica Colloidal Crystals

被引:35
作者
Wei, Bingchuan [1 ]
Malkin, Douglas S. [1 ]
Wirth, Mary J. [1 ]
机构
[1] Purdue Univ, Dept Chem, W Lafayette, IN 47907 USA
关键词
PERFORMANCE LIQUID-CHROMATOGRAPHY; CAPILLARY-ZONE-ELECTROPHORESIS; MASS-SPECTROMETRY; SEPARATION; PRESSURE; PARTICLES; PHASE; COLUMNS; SPEED; PROTEOMICS;
D O I
10.1021/ac102438w
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
Silica colloidal crystals formed from 330 nm nonporous silica spheres inside of 75 mu m i.d. fused silica capillaries were evaluated for the efficiency of capillary electrochromatography of proteins. Three proteins, ribonuclease A, cytochrome C, and lysozyme, each covalently labeled with fluorophor, were well separated over a distance of 1 cm by isocratic electromigration, using 40:60 acetonitrile/water with 0.1% formic acid. A van Deemter plot showed that the plate height for lysozyme, which was the purest of the three proteins, was diffusion-limited for electric fields ranging from 400 to 1400 V/cm. The plate height for lysozyme was below 50 nm at almost all of the migration velocities, and it approached 10 nm at the highest velocity. Eddy diffusion was negligible. Lysozyme migrated over a 12 mm separation length with more than 10(6) plates in 1.5 min. These results indicate that silica colloidal crystals are well suited for electrically driven separations of large, highly charged analytes such as proteins. The 10(6) plates observed for a separation length of barely more than a centimeter means they are potentially valuable for miniaturized separations in microchip and lab-on-a-chip devices.
引用
收藏
页码:10216 / 10221
页数:6
相关论文
共 35 条
  • [1] Proteomics by FTICR mass spectrometry: Top down and bottom up
    Bogdanov, B
    Smith, RD
    [J]. MASS SPECTROMETRY REVIEWS, 2005, 24 (02) : 168 - 200
  • [2] PREDICTING PROTEIN DIFFUSION-COEFFICIENTS
    BRUNE, D
    KIM, S
    [J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (09) : 3835 - 3839
  • [3] SilicaROD™ -: A new challenge in fast high-performance liquid chromatography separations
    Cabrera, K
    Wieland, G
    Lubda, D
    Nakanishi, K
    Soga, N
    Minakuchi, H
    Unger, KK
    [J]. TRAC-TRENDS IN ANALYTICAL CHEMISTRY, 1998, 17 (01) : 50 - 53
  • [4] Effect of Pressure, Particle Size, and Time on Optimizing Performance in Liquid Chromatography
    Carr, Peter W.
    Wang, Xiaoli
    Stoll, Dwight R.
    [J]. ANALYTICAL CHEMISTRY, 2009, 81 (13) : 5342 - 5353
  • [5] Future of high pressure liquid chromatography: Do we need porosity or do we need pressure?
    Desmet, Gert
    Cabooter, Deirdre
    Gzil, Piotr
    Verelst, Harry
    Mangelings, Debby
    Vander Heyden, Yvan
    Clicq, David
    [J]. JOURNAL OF CHROMATOGRAPHY A, 2006, 1130 (01) : 158 - 166
  • [6] Characteristics of superficially-porous silica particles for fast HPLC:: Some performance comparisons with sub-2-μm particles
    DeStefano, J. J.
    Langlois, T. J.
    Kirkland, J. J.
    [J]. JOURNAL OF CHROMATOGRAPHIC SCIENCE, 2008, 46 (03) : 254 - 260
  • [7] Ding WL, 1997, HRC-J HIGH RES CHROM, V20, P575
  • [8] Euerby MR, 1997, J MICROCOLUMN SEP, V9, P373
  • [9] USE OF METHYL SPACERS IN A MIXED HORIZONTALLY POLYMERIZED STATIONARY-PHASE
    FAIRBANK, RWP
    XIANG, Y
    WIRTH, MJ
    [J]. ANALYTICAL CHEMISTRY, 1995, 67 (21) : 3879 - 3885
  • [10] CAPILLARY ZONE ELECTROPHORESIS SEPARATIONS OF BASIC AND ACIDIC PROTEINS USING POLY(VINYL ALCOHOL) COATINGS IN FUSED-SILICA CAPILLARIES
    GILGES, M
    KLEEMISS, MH
    SCHOMBURG, G
    [J]. ANALYTICAL CHEMISTRY, 1994, 66 (13) : 2038 - 2046